Composite mutagenesis and protoplast fusion have been employed in an attempt to improve the production of S-adenosyl-L-methionine (SAM) by S. cerevisiae R1 The haploidy of R1, hR10, was prepared by enzymatic hydrolysis and treated with UV and diethylstilbesterol (DES). Four haploid mutants named DR1-3, NM14, NM39, and NM45 were selected and their protoplasts were grouped. One group was inactivated by heat, and another group was inactivated by UV irradiation. Then polyethylene glycol (PEG) was used to mediate protoplast fusion and one fusant F35 with improved production of SAM was obtained by screening. The production of SAM by F35 reached 22.5 mg/L, an increase of 103% compared with the value for the starting strain R1 Subculture experiments indicated that the hereditary character of high production was stable. The results of orthogonal tests showed that a medium containing 100 g/L of maltose, 10 g/L of peptone, 4g/L of NHCH2PO4, 5 g/L of NH4Cl, 0.1 g/L of MgSO4, and 6 g/L of KH2PO4 with a fermentation time of 72 h was suitable for production of SAM by F35.
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